标题:High efficient transformation of auxin reporter gene into trifoliate orange via Agrobacterium rhizogenes-mediated co-transformation

作者:Xiao, X., Ma, F., Chen, C., and Guo, W

摘要:An efficient Agrobacterium rhizogenes mediated transformation system of Poncirus trifoliata was developed. The combination of A. rhizogenes MSU440 with epicotyl segments was confirmed to be02a suitable system for genetic transformation in P. trifoliata . About 302weeks after inoculation with bacteria, hairy roots emerged at the cut surfaces of epicotyl segments on Murashige and Tucker selective medium free of plant growth regulators. The presence of rol B, β-glucuronidase (GUS) and yellow fluorescent protein (YFP) genes in co-transformed hairy roots was confirmed by fluorescence observation, GUS staining and polymerase chain reaction (PCR) analysis. The co-transformation rate based on GUS staining was as high as 68.302%. A regeneration procedure of transgenic plantlets from hairy roots was also established. Out of 45 hairy root segments from different co-transformed lines, 23 were successfully regenerated into rooted plantlets. The presence and expression of YFP gene in the regenerated co-transformed plants were further confirmed by Southern blot hybridization and quantitative reverse transcription PCR. The DR5 ∷ GUS – YFP reporter system was used to study the auxin response maxima at the apices of primary hairy roots and their lateral roots. Auxin distribution and response maxima during lateral root primordium development were also studied. The hairy roots obtained by A. rhizogenes mediated transformation were confirmed to be a good system for investigating root development in P. trifoliata . Furthermore, the genetic transformation and plant regeneration system via A. rhizogenes provides an alternative method for genetic engineering in Citrus .

关键词:Poncirus trifoliata ,Agrobacterium rhizogenes ,Hairy root , DR5 ,Plant regenerat

文献注录:Xiao, X., Ma, F., Chen, C., and Guo, W. High efficient transformation of auxin reporter gene into trifoliate orange via Agrobacterium rhizogenes-mediated co-transformation [J]. Plant Cell Tissue & Organ Culture. 2014, 118(1): 137-146.

报/刊名:Plant Cell Tissue & Organ Culture》,发表于2014 / 第 118(1) 期。

文献类型: [J]

页码: 137-146 页 / 共10

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